ES2298497T3 - QUINASA RHO INHIBITORS. - Google Patents

Abstract

A compound of the formula (See formula)

Description

Rho kinase inhibitors.

Field of the Invention

The present invention relates to compounds and derivatives thereof, their synthesis, and their use as inhibitors of Rho kinase. These compounds of the present invention are useful. to inhibit tumor growth, treat erectile dysfunction, and treat other indications mediated by Rho kinase, by example, coronary heart disease.

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Background

The pathology of a series of diseases humans and animals including hypertension, dysfunction erectile, coronary cerebral circulatory impairments, neurodegenerative disorders and cancer can be linked directly to changes in the actin cytoskeleton. These Diseases pose a serious unmet medical need. He Actin cytoskeleton is composed of a filament mesh of actin and actin-binding proteins found in All eukaryotic cells. In the muscles cells Smooth assembly and disassembly of the actin cytoskeleton is the main driving force responsible for the contraction and the relaxation of smooth muscles. In non-muscle cells, the Dynamic transpositions of the actin cytoskeleton are responsible for the regulation of cell morphology, motility  cellular, the formation of actin tensile fibers, the cell adhesion and specialized cell functions such such as neurite retraction, phagocytosis or cytokinesis (Van Aelst, et al., Genes Dev 1997, 11, 2295).

The actin cytoskeleton is controlled by a family of proteins that are a subset of the Ras superfamily of GTPases. This subset is constituted currently by RhoA to E and RhoG (collectively referred to as Rho), Rac 1 and 2, Cdc42Hs and the G25K and TC10 isoforms (Mackay, and col., J. Biol. Chem. 1998, 273, 20685). These proteins are proteins that bind to GTP (guanine nucleotide triphosphate) with intrinsic GTPase activity. They act as switches and cycles molecules between the United States to GDP (nucleotide diphosphate guanine) inactive and linked to active GTP. Using biochemical and genetic manipulations, it has been possible to assign functions to each family member. After activation the Rho proteins control the formation of tensile fibers of actin, thick bundles of actin filaments, and the grouping of integrins in focal adhesion complexes. When activated Rac proteins control the formation of lamelipodia or waves of membrane on the cell surface and Cdc42 controls the formation of phyllopods Together this family of proteins plays a critical part in the control of key cellular functions including cell movement, axon guidance, cytokinesis, and changes in cell morphology, shape and polarity.

Depending on the type of cell and receptor that is activated, Rho proteins can control different biological responses In the smooth muscle cells, the Rho proteins are responsible for calcium sensitization during the contraction of smooth muscles. In cells that are not of the smooth muscles Rho GTPases are responsible for the cellular responses to agonists such as lysophosphatidic acid (LPA), thrombin and thromboxane A2 (Fukata, et al. Trends Pharcol Sci 2001, 22, 32). The response to the agonist is coupled to through the heterotrimeric G alpha12 proteins or G_13 (Goetzl, et al. Cancer Res 1999, 59, 4732; Buhl, and cabbage. J. Biol. Chem. 1995, 270, 24631) although they may be involved other receivers Upon activation Rho GTPases activate some down-chain effectors including PIP5 kinase, the Rhotequina, Rhofilina, PKN and Rho kinase isoforms ROCK-1 / ROKbeta and ROCK-1 / ROKalfa (Mackay and Hall J. Biol. Chem. 1988, 273, 20685; Aspenstrom Curr Opin Cell Biol 1999, 11, 95; Amano et al. Exp. Cell Res 2000, 261, 44).

Rho kinase was identified as a protein that interacts with RhoA isolated from the bovine brain (Matsui, et al. Embo J 1996, 15, 2208). He is a member of the family of myotonic protein kinase dystrophy and contains a serine / threonine kinase domain at the amino terminus, a domain spirally wound in the central region and a domain of interaction with Rho at the carboxy terminus (Amano et al. Exp. Cell Res 2000, 261, 44). Its kinase activity is increased after binding to RhoA bound to GTP and when it is introduced into cells, it can reproduce many of the activated RhoA activities. In the smooth muscle cells kinase Rho mediates the calcium sensitization and smooth muscle contraction and Rho kinase inhibition blocks muscle contraction induced by 5-HT and phenylephrine agonists. When it is introduced into cells that are not of the smooth muscles, the Rho kinase induces the formation of tensile fibers and is required  for RhoA-mediated cell transformation (Sahai, et al. Curr Biol 1999, 9, 136). Rho kinase regulates a series of proteins chain down through phosphorylation, including chain light myosin (Somlyo, et al. J Physiol (Lond) 2000, 522 Pt 2, 177), the subunit that binds to the light chain phosphatase of myosin (Fukata, et al. J. Cell Biol 1998, 141, 409) and the kinase 2 LIM (Sumi, et al., J. Bio Chem 2001, 276, 670).

Inhibition of Rho kinase activity in animal models has demonstrated a number of benefits of Rho kinase inhibitors for the treatment of human diseases. Several patents have appeared claiming compounds of (+) - trans-4- (1-aminoethyl) -1- (pyridin-4-ylaminocarbonyl) cyclohexane dihydrochloride (WO-00078351, WO-00057913) and substituted isoquinolinosulfonyl compounds (document EP-00187371) as Rho kinase inhibitors with activity in animal models. These include models of cardiovascular diseases such as hypertension (Uehata, et al. Nature 1997, 389, 990), atherosclerosis (Retzer, et al. FEBS Lett. 2000, 466, 70), restenosis (Eto, and col. Am J. Physiol Heart Circ Physiol 2000, 278, H1744; Negoro, et al. Biochem Biophys Res Commun 1999, 262, 211), cerebral ischemia (Uehata, et al. Nature 1997, 389, 990: Seasholtz et al. ., Circ Res 1999, 84, 1186; Hitomi, et al. Life Sci 2000, 67, 1929; Yamamoto, et al. J Cardiovasc Pharmacol 2000, 35, 203), cerebral vasospasm (Sato, et al, Circ Res 2000 , 87, 195; Kim, et al. Neurosurgery 2000, 46, 440), penile erectile dysfunction (Chitaley, et al. Nat Med 2001, 7, 119), central nervous system disorders such as neuronal degeneration and spinal cord injury (Hara, et al. J Neurosurg 2000, 93, 94; Toshima, et al. Stroke 2000, 31, 2245) and in neoplasms where Rho kinase inhibition has been shown to inhibit growth to tumor cells and metastasis (Itoh, et al. Nat Med 1999, 5, 221; Somlyo, et al. Biochem Biophys Res Commun 2000, 269, 652), angiogenesis (Uchida, et al., Biochem Biophys Res Commun 2000, 269, 633; Gingras, et al. Biochem J 2000, 348 Pt 2, 273), arterial thrombotic disorders such as platelet aggregation (Klages, et al. J. Cell Biol 1999, 144, 745; Retzer et al. Cell Signal 2000, 12, 645) and leukocyte aggregation (Kawaguchi, et al., Eur J Pharmacol 2000 , 403, 203; Sánchez-Madrid, et al., Embo J 1999, 18, 501), asthma (Setoguchi, et al. Br J Pharmacol 2001, 132, 111; Nakahara, et al. Eur J Pharmacol 2000, 389 , 103), the regulation of intraocular pressure (Honjo, et al. Invest Ophthalmol Vis Sci 2001, 42, 137) and bone resorption (Chellaiah, et al. J Biol Chem 2000, 275, 11993; Zhang, et al. J Cell Sci 1995, 108,
2285).

Inhibition of Rho kinase activity in patients it has benefits to control vasospasms brain and ischemia following subarachnoid hemorrhage (Pharma Japan 1995, 1470, 16).

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Summary of the invention

The compounds and their derivatives presented in this invention are useful as Rho kinase inhibitors and by both have utilities in the treatment of hypertension, the atherosclerosis, restenosis, cerebral ischemia, vasospasm cerebral, neuronal degeneration, spinal cord injury, cancers of the breast, colon, prostate, ovaries, brain and lung and their metastases, thrombotic disorders, asthma, glaucoma and osteoporosis

In addition, the compounds of the invention are useful for treating erectile dysfunction, that is, dysfunction erectile mediated by Rho kinase. Erectile dysfunction can be defined as an inability to obtain or maintain an erection suitable for intercourse, documents WO 94/28902, U.S.P. 6,103,765 and U.S.P. 6,124,461.

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The invention involves compounds of the following structures:

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one

2

3

4

5

6

7

8

9

10

eleven

12

13

14

The compounds of Formula I can be elaborate according to conventional chemical procedures, routine, and / or as described below, from materials which are either available in the market or Producible according to conventional chemical procedures, routine The procedures for the preparation of Compounds are given below in the Examples.

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In the following examples, all temperatures are exposed uncorrected in degrees Celsius; and unless otherwise indicated, all parts and percentages are in weight.

The full description of all applications, patents and publications, cited above or then, and Provisional Application No. 60 / 349,986, filed on January 23, 2002, they are incorporated herein by reference.

Abbreviations and acronyms

When the following abbreviations are used in the present specification, they have the following meaning:

Ac_ {2} O

acetic anhydride

anhi

anhydrous

n-BuOH

n-butanol

t-BuOH

t-butanol

CD_ {3} OD

methanol-d4

Celite®

diatomaceous earth filtration agent, ® Celite Corp.

CH 2 Cl 2

methylene chloride

CI-MS

ionization mass spectroscopy chemistry

conc

concentrated

desc

decomposition

DME

dimethoxyethane

DMF

N, N-dimethylformamide

DMSO

dimethylsulfoxide

ELSD

evaporative dispersion detector light

EtOAc

ethyl acetate

EtOH

ethanol (100%)

Et 2 O

diethyl ether

Et 3 N

triethylamine

HPLC ES-MS

chromatography High resolution liquid mass spectroscopy electrospray

NMM

4-methylmorpholine

Ph_ {3} P

triphenylphosphine

Pd (dppf) Cl_ {2}

[1,1'-bis (diphenylphosphino) ferrocene] -dichloropaladium (II)

Pd (PPh 3) 4

tetrakis (triphenylphosphine) palladium (0)

Pd (OA_c) 2

palladium acetate

P (O) Cl3   

phosphorus oxychloride

RT

retention time (HPLC)

rt

room temperature

THF

tetrahydrofuran

TFA

trifluoroacetic acid

TCL

Thin layer chromatography

Experimental examples

All reactions were performed in glass flame drying or oven drying under a positive pressure of dry argon, and stirred magnetically unless indicated another thing. Sensitive liquids and solutions were transferred to through a syringe or cannula, and were introduced into the vessels of the reaction through rubber septa. The reagents and solvents of Commercial grade were used without further purification. The Thin layer chromatography (TLC) was performed on 250 µm plates pre-coated with silica gel 60 A F-254 reinforced with glass from Analtech UNIPLATE?. Chromatography Column (ultra-fast chromatography) was performed in a Biotage system using pre-packaged cartridges silica gel, 60 A, 32-63 micrometers. Resonance Proton nuclear magnetic (1 H) (NMR) was measured with a Varian spectrometer (300 MHz) with residual protonated solvent (CHCl 3 δ 7.26; MeOH δ 3.30; DMSO δ 2.49) as a patron Low resolution mass spectra (MS) are obtained either as mass spectra by electronic impact (IE) or as mass spectra by rapid atomic bombardment (PUB).

The IUPAC designation was obtained using the ACD / ILab Web service.

A. Preparation of chloropyrimidine intermediates

Intermediate A1

Preparation of 2-amino-4-chloro-5,6-dimethyl-pyrimidine

fifteen

Stage one

Preparation of 2-amino-5,6-dimethyl-pyrimidinone

16

To a solution of 2-acetoacetate ethyl (6.0 g, 41.6 mmol) and guanidine carbonate (5.6 g, 31.2 mmol) in EtOH (32 ml) 12 N HCl (350 µl) was added. The mixture is kept at reflux for 16 hours. Then the reaction cooled. at room temperature, the solid was collected by filtration and was washed with EtOH. A solution of the solid was refluxed in NaOH 1 N for 3 hours. Then, the reaction was cooled to temperature. ambient, the aqueous mixture was adjusted to pH = 5 with acetic acid concentrated. The resulting precipitate was collected by filtration, washed with water and then with hexanes, and dried under vacuum. He desired compound (6.34 g, 45.6 mmol; 100% yield); 1 H NMR (D 2 O; NaOD) δ 1.47 (s, 3H), 1.29-1.30 (m, 2H), 1.22 (s, 3 H); ES MS [M + H] + = 140.

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Stage 2

Preparation of 2-amino-4-chloro-5,6-dimethyl-pyrimidine

17

The product of the previous stage (2.0 g, 14.4 mmol) and phosphorus oxychloride (6 ml, 57.5 mmol), was maintained at reflux for 4 hours. The reaction was cooled to rt and poured on ice. The mixture was separated and the aqueous phase was extracted with chloroform (3 x 75 ml). The aqueous mixture was adjusted to pH = 9 with concentrated ammonium hydroxide. The resulting solid product is collected by filtration, washed with water, and dried in vacuo. He desired compound (963 mg, 6.1 mmol; 43% yield); mp = 212 - 220 ° C; ES MS [M + H] + = 158; TLC (CH 2 Cl 2 - MeOH, 90:10); R f = 0.72.

Intermediate A2

Preparation of 2-amino-4-chloro-6- (4-pyridyl) pyrimidine

18

Stage one

Preparation of 2-amino-4-hydroxy-6- (4-pyridyl) pyrimidine

19

A solution of guanidine carbonate (7.1 g, 39 mmol, 1.5 eq), ethyl isonicotinoyl acetate (10 g, 51.76 mmol), and hydrochloric acid (0.75 ml, 9.0 mmol) in Absolute ethanol (80 ml) was refluxed under argon overnight. The precipitate formed was filtered, washed with ethanol and dried. The solid was then dissolved in 1 N NaOH (100 ml) and refluxed for 2 hours. The reaction mixture was then cooled to room temperature, acidified with glacial acetic acid, and the solid formed was filtered and dried to provide the desired product as a white solid (5.45 g, 56%). 1 H NMR (DMSO- δ 6) δ 6.24 (s, 1H), 6.79 (sa, 2H), 7.85 (d, J = 5.1 Hz, 2H ), 8.62 (d, J = 5.3 Hz, 2H), 11.22 (sa, 1H).

Stage 2

Preparation of 2-amino-4-chloro-6- (4-pyridyl) pyrimidine

twenty

A solution of 2-amino-4-hydroxy-6- (4-pyridyl) pyrimidine (5.45 g, 29 mmol) in POCl 3 (12 ml) was refluxed under argon for 5 hours. The reaction mixture was cooled to room temperature, poured onto ice, and allowed to stir at room temperature for 2 hours to ensure the interruption of POCl3. At that time, the mixture became basic after the addition of 1 N NaOH and the brown solid was filtered to provide 4.52 g of crude product, which was used without further purification (NMR analysis showed 1: 1 product / material of departure). The filtrate formed more solid after remaining at room temperature (1 g, NMR analysis showed 2: 1 product / starting material). 1 H NMR (DMSO- δ 6) δ 7.34 (sa, 2H), 7.38 (s, 1H), 7.99 (d, J = 4.2 Hz, 2H ), 8.72 (d, J = 4.6 Hz, 2H).

Intermediate A3

Preparation of 2-amino-4-chloro-6- (2-thienyl) pyrimidine

twenty-one

Stage one

Preparation of ethyl-2- (thiophene-2-oil) acetate

22

A solution of thiophene-2-carboxylic acid (8.9 g, 68.5 mmol), 2,2-dimethyl-1,3-dioxane-4,6-dione (12.0 g, 81.6 mmol), and 4-dimethylaminopyridine (17.0 g, 138 mmol) in dry CH 2 Cl 2 (100 ml) was cooled to 0 ° C and treated with a solution of 1,3-dicyclohexylcarbodiimide (75 ml, 1.0 M in CH 2 Cl 2, 75 mmol). The reaction was allowed to stir at room temperature for 2 hours and the dicyclohexylurea was then filtered and washed with CH2Cl2. The filtrate was concentrated under reduced pressure and the residue was dissolved in absolute ethanol (400 ml). The solution was then treated with a solution of p- toluenesulfonic acid monohidate (32 g, 168 mmol) in absolute ethanol (100 ml) and refluxed under argon for 1 hour. At that time, the ethanol was removed under reduced pressure and the residue was dissolved in EtOH and washed sequentially with H 2 O (300 ml), saturated NaHCO 3 (200 ml), 1 N HCl (200 ml) , Saturated NaCl, and dried (MgSO 4). The solvent was removed under reduced pressure and the residue was filtered through a bed of silica with 10% EtOAc / 90% hexanes to provide the desired product as an oil (13 g, 96%). TLC (20% EtOAc / 80% hexane) R f 0.21; 1 H NMR (DMSO- d 6) δ 1.17 (t, J = 7.01, 3H), 4.06-4.14 (m, 4H), 7.25 (t , J = 5.1 Hz, 1H), 7.98 (d, J = 3.8 Hz, 1H), 8.06 (d, J = 4.9 Hz, 1H).

Stage 2

Preparation of 2-amino-4-hydroxy-6- (2-thienyl) pyrimidine

2. 3

The procedure was similar to that used for Intermediate A2, step 1, using ethyl-2- (thienyl-2-oil) acetate as the starting material. (Yield at 43%). TLC (6% MeOH / 94% CH 2 Cl 2) R f 0.23; MS ES 194 [M + H] +; 1 H NMR (DMSO- d 6) δ 6.06 (s, 1H), 6.70 (sa, 2H), 7.11 (t, J = 4.9 Hz, 1H) , 7.64 (d, J = 4.9 Hz, 1H), 7.70 (d, J = 3.6 Hz, 1H), 10.95 (sa, 1H).

Stage 3

Preparation of 2-amino-4-chloro-6- (2-thienyl) pyrimidine

24

The procedure was similar to that used for Intermediate A2, step 2, using 2-amino-4-hydroxy-6- (2-thiophene) pyrimidine as the starting material. (Provided a 33% yield after purification on silica with 15% EtOAc / 85% hexanes. TLC (20% EtOAc / 80% hexanes) R f 0.29; 1 H NMR (DMSO- d 6) δ 7.16-7.23 (m, 4H), 7.77 (dd, J = 0.8, 5.0 Hz, 1H), 7.98 (dd , J = 1.0, 3.8 Hz, 1H).

Intermediate A4

Preparation of 2-amino-4-chloro-6- (2-furyl) pyrimidine

25

Stage one

Preparation of 2-amino-4-hydroxy-6- (2-furyl) pyrimidine

26

The general procedure was used for preparation of Intermediate A2, (stage 1); (37% yield). MS (ES) 178 [M + H] +.

Stage 2

Preparation of 2-amino-4-chloro-6- (2-furyl) pyrimidine

27

A solution of 2-amino-4-hydroxy-6- (2-furyl) pyrimidine (1.40 g, 7.9 mmol) in POCl 3 (4 ml) was refluxed under argon for 2 hours. The POCl3 was distilled; The residue was diluted with EtOAc and poured onto ice cold saturated NaHCO3. The phases were separated and the aqueous one was extracted with EtOAc (100 ml). The combined extracts were washed with saturated NaCl, dried (MgSO 4), and the solvent was removed under reduced pressure to provide 0.5 g of crude product, which was used without further purification. TLC (20% EtOAc / 80% hexane) R f 0.26; 1 H NMR (DMSO-d 6) δ 6.68 (dd, J = 1.7.3.4 Hz, 1H), 6.94 (s, 1H), 7.25 (dd , J = 1,3.7 Hz, 1H), 7.91 (dd, J = 0.8, 1.9 Hz, 1H).

Intermediate TO 5

Preparation of 6-benzyl-4-chloro-5,6,7,8-tetrahydropyrid [4,3- d ] pyrimidin-2-amine

28

Stage one

Preparation of 2-amino-7-benzyl-5,6,7,8-tetrahydropyrid [3,4- d ] pyrimidin-4 (3H) -one

29

To EtOH (16 ml) cooled to 0 ° C (ice bath / H2O) Na spheres (204 mg, 8.9 mmol) were added. The mixture is stirred until all Na dissolved. Hydrochloride was added 1-Benzyl-4-oxo-3-piperidine carboxylate  methyl (3.0 g, 10.1 mmol) and guanidine carbonate (1.4 g, 7.6 mmol). The mixture was refluxed for 16 hours. After the reaction was cooled to room temperature, the solid was collected by filtration, washed with EtOH, and dried in vacuo. The compound desired (2.58 g, 10.0 mmol; 99 +% yield); mp = 202 - 212º (dec.); ES MS [M + H] + = 257; TLC (CH 2 Cl 2 - MeOH, 90:10); R_ {f} = 0.20.

Stage 2

Preparation of 6-benzyl-4-chloro-5,6,7,8-tetrahydropyrid [4,3- d ] pyrimidin-2-amine

30

A solution of the product of step 1 (3.5 g, 13.7 mmol) in POCl3 (52 ml) was refluxed under argon for 5 hours The reaction mixture was cooled to temperature. ambient, poured on ice, and allowed to stir at temperature environment for 2 hours to ensure the interruption of POCl_ {3}. At that time, the mix became basic after the addition of ammonium hydroxide and extracted with CH2Cl2 (3 x 200 ml). The combined organic extracts were washed with 1 N NaOH followed by brine, dried (MgSO4), and concentrated to reduced pressure The residue was taken up in benzene and made acidic. after the addition of 1 N HCl in diethyl ether. The brown solid is filtered to provide 0.35 g of crude product, which was used without additional purification ES MS [M + H] + = 275.

Intermediate A6

Preparation of 2-amino-6- (trifluoromethyl) -4-pyrimidinyl-4-methylbenzenesulfonate

31

To a solution of 2-amino-6- (trifluoromethyl) -4 (3 H -pyrimidinone) (250 mg, 1.4 mmol), triethylamine (196 µL, 1.4 mmol), N, N dimethylaminopyridine (17 mg , 0.14 mmol), in CH 2 Cl 2 (13 ml) cooled to 0 ° C. p-toluenesulfonyl chloride (534 mg, 2.8 mmol) was added. The mixture was stirred at room temperature for 16 h. The mixture was diluted with CH2Cl2, washed with H2O (2 x 20 mL) followed by brine, dried (Na2SO4), evaporated, and evaporated. Vacuum dried. The desired compound (466 mg, 1.4 mmol; 99 +% yield; ES MS [M + H] + = 140.

Using the above procedures for preparation of A1-A6 and replacing materials appropriate starting materials, the following were also prepared pyrimidine intermediates.

TABLE 1

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B. Preparation of arylamine intermediates

Intermediate B1

Preparation of 1- (4-pyridinyl) -1 H -indole-5-amine

3. 4

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Stage one

Preparation of 5-nitro-1- (4-pyridinyl) -1 H -indole

35

To a solution of 5-nitroindole (7.0 g, 43.2 mmol) and 4-chloropyridine hydrochloride (7.8 g, 51.8 mmol) in DMF (43 ml) was added potassium tert-butoxide (12 , 1 g, 108.0 mmol), in portions. The reaction was heated at 100 ° C for 48 hours. The mixture was allowed to cool to room temperature and poured into water (400 ml). The resulting solid was filtered off and dried in vacuo. The desired compound (6.04 g, 25.3 mmol; 58% yield); 1 H NMR (DMSO- d 6) δ 8.76 (dd, J = 1.7, 4.5, 2H), 8.68 (d, J = 2.2, 1H), 8.06 - 8.13 (m, 2H), 7.92 (d, J = 9.2, 1H), 7.75 (dd, J = 1.5, 4.6, 2H), 7.07 (dd, J = 0.9, 3.5, 1H); ES MS [M + H] + = 240.

Stage 2

Preparation of 1- (4-pyridinyl) -1 H -indole-5-amine

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36

A mixture of the product of step 1 (8.27 g, 34.6 mmol) and 10% palladium on charcoal catalyst (827 mg) in ethyl acetate (166 ml) and EtOH (9 ml) was stirred in hydrogen at atmospheric pressure for 48 hours. Another was added catalyst (414 mg) and the reaction was stirred for 24 hours. From again, another catalyst (414 mg) was added and the reaction was stirred 24 hours more. The catalyst was removed by filtration through diatomaceous earth and the solvent was removed from the filtrate by evaporation. The residue was triturated with ether, collected by filtration, and dried in vacuo. The desired compound (4.67 g, 22.3 mmol; 65% yield); mp = 149-154; ES MS [M + H] + = 210; TLC (CH 2 Cl 2 -MeOH, 95: 5); R f = 0.29.

Intermediate B2

Preparation of 4 - [(4-aminophenyl) sulfanyl] phenol

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37

Stage one

Preparation of 4 - [(4-nitrophenyl) sulfanyl] phenol

38

To a chloride solution of nitrobenzenesulfonyl (4 g, 21 mmol) in ether (25 ml) phenol was added (1.97 g, 20 mmol) as a solution in ether (25 ml). After Stirring for 15 hours at rt, the mixture was concentrated to provide a crude solid that was recrystallized from acid acetic.

The desired compound (4.0 g, 16.2 mmol, 76% yield). TLC (Hexanes / EtOAc, 70:30); R_ {f} = 0.54.

Stage 2

Preparation of 4 - [(4-aminophenyl) sulfanyl] phenol

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39

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To a solution of the product of step 1 (4 g, 16.2 mmol) in EtOH (500 ml) SnCl 2 • 2H 2 O was added (18.3 g, 81 mmol). The solution was heated to reflux. After Stir for 3 hours, the mixture was allowed to cool to rt, and the Volatile compounds were removed by rotary evaporator. The suspension The resulting was suspended in EtOAc, and solid NaHCO3 was added. Subsequently, the mixture was filtered and the filtered solid washed conscientiously with EtOAc. The organic filtrate was washed with water, and the aqueous washings were extracted with EtOAc. Extracts The combined organics were washed with brine, dried (MgSO4), filtered and concentrated to provide an orange solid, which was used without further purification. Desired compound (3.0 g, 13.8 mmol, 86% yield).

TLC (Hexanes / EtOAc, 70:30); R_ {f} = 0.34.

Intermediate B3

Preparation of (3-aminophenyl) [4-methylsulfanyl) phenyl] methanone

40

Stage one

Preparation of [4- (methylsulfanyl) phenyl] (3-nitrophenyl) methanone

41

3-Nitrobenzoylchloride (5.0 g, 26.94 mmol) was added to a solution of thioanisole (3.16 ml, 26.94 mmol) and 1,2-dichloroethane (95 ml). The resulting reaction mixture was cooled to 0 ° C (ice bath / H2O) and 0.5 equivalents of aluminum trichloride (1.8 g, 13.47 mmol) was added. The reaction was allowed to stir for 15 minutes at this temperature and the cold bath was removed followed by the addition of the remaining equivalents of AlCl 3 (2.51 g, 18.87). The reaction solution turned a dark yellow / greenish and was allowed to stir at room temperature for 18 hours, after which time the reaction was slowly stopped with H2O (50 ml). The mixture was diluted with CH2Cl2 (50 ml) and washed with H2O (3 x 50 ml), and the combined organic phases were washed with saturated NaHCO3 (50 ml) , dried (MgSO 4) and concentrated under reduced pressure. The crude material was purified by column chromatography on silica gel (EtOAc / hexane, ¼) to provide 3.3 g (44%) of 4- (methylsulfanyl) phenyl] (3-nitrophenyl) methanone as a solid. EI-LRMS m / z 274 (M +); TLC R f 0.68 (EtOAc / Hex, 2/3).

Stage 2

Preparation of (3-aminophenyl) [4- (methylsulfanyl) phenyl] methanone

42

Prepared analogously to Intermediate B2, step 2. The crude product was purified by column chromatography. ultra-fast, eluting with 70:30 Hexanes / EtOAc. TLC: (Hexanes / EtOAc, 70:30);

R f = 0.15.

Intermediate B4

Preparation of 4- (4-aminophenoxy) phenol

43

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Stage one

Preparation of 4- (4-nitrophenoxy) phenol

44

A mixture of p- nitrofluorobenzene (25 g, 0.177 mol), dihydroquinone (19.5 g, 0.177 mol), and sodium hydroxide (7.08 g, 0.177 mol) in EtOH / H2O (1: 1 v / v, 176 ml) was heated at reflux for 20 hours, and subsequently allowed to cool to room temperature. The mixture was filtered, the filtrate was made acidic with dilute aqueous HCl, and the resulting precipitate was filtered to give the crude product as a yellow solid. The desired product was recrystallized from EtOH. (15 g, 0.064 mol, 37% yield). TLC (Hexanes / EtOAc, 70:30); R_ {f} = 0.44.

Stage 2

Preparation of 4- (4-aminophenoxy) phenol

Four. Five

To a solution of the product of stage 1 in EtOH (100 ml) 10% palladium on carbon (200 mg) was added. After stirring under a hydrogen atmosphere throughout the entire night, the mixture was filtered through Celite ®. The compounds volatiles were removed from the filtrate to provide the product crude that was purified by column chromatography ultra-fast eluting with Hexanes / EtOAc (85:15, followed by 75:25). Desired product (1.5 g, 7.45 mmol; 86%). FTA (Hexanes / EtOAc, 70:30); R_ {f} = 0.41.

Intermediate B5

Preparation of 4- (4-pyridinylthio) aniline

46

To a solution of 4-aminothiophenol (20.2 g, 156.5 mmol) in anhydrous DMF (200 ml) was added 4-chloropyridine hydrochloride (24.4 g, 161.0 mmol) followed by potassium carbonate (44 g, 318.4 mmol). The reaction mixture was heated at 80 ° C overnight, then ethyl acetate (400 ml) and water (400 ml) were diluted. The aqueous phase was reextracted with ethyl acetate (2 x 200 ml). The combined organic phases were washed with a saturated aqueous solution of NaCl (200 ml), dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was filtered through a bed of silica with ethyl acetate and the resulting material was triturated with an ethyl ether / hexane solution to provide the desired product (24.7 g, 78%). TLC (50% ethyl acetate / 50% hexane) R f 0.25; 1 H NMR (DMSO-d 6) δ 5.67 (sa, 2H), 6.65 (d, J = 8.4 Hz, 2H), 6.88 (d, J = 6 , 2 Hz, 2H), 7.19 (d, J = 8.4 Hz, 2H), 8.27 (d, J = 6, 2 Hz, 2H), MS [M + H] + = 203

Intermediate B6

Preparation of 4- [2- (4-pyridinyl) ethyl] aniline

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Stage one

Preparation of 4 - [( E ) -2- (4-nitrophenyl) ethenyl] pyridine

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To a 500 ml 3-mouth flask dried in the oven bromide of (4-nitrobenzyl) triphenylphosphonium (15 g, 30.42 mmol) followed by the addition of THF (100 ml). The solution was cooled to 0 ° C in an ice bath. Then it was added potassium t-butoxide (3.9 g, 33.02 mmol) in one portion resulting in an orange suspension. The suspension is held at 0 ° C while adding a solution of 4-pyridine-2-carboxaldehyde (2.7 g, 24.70 mmol) in THF (20 ml) in 10 minutes. Ice bath it was removed and the reaction was stirred at room temperature for 2 hours. At that time the reaction was stopped with solution of saturated ammonium chloride (50 ml) and stirred for 15 minutes. After the mixture was extracted with ethyl acetate (2 x 100 ml), the The combined extracts were washed with saturated aqueous NaCL solution. (100 ml) and dried (MgSO 4). The solvent was removed at reduced pressure and the residue was chromatographed on silica with 0% -50% ethyl acetate in hexanes to provide the desired product (1.8 g, 32%). TLC (50% ethyl acetate / 50% hexane) R f 0.28; 1 H NMR (DMSO-d6) δ 6.84 (d, J = 12.4Hz, 1H), 6.96 (d, J = 12.4Hz, 1H), 7.14 (d, J = 6.2Hz, 2H), 7.45 (d, J = 8.7Hz, H), 8.15 (d, J = 8.7Hz, 2H), 8.47 (d, J = 6.2Hz, 2H).

Stage 2

Preparation of 4- [2- (4-pyridinyl) ethyl] aniline

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To a dry 50 ml flask purged with argon Pd on 10% carbon (285 mg) was added followed by the addition of ethanol (12 ml) and the product of step 1 (1.8 g, 8.0 mmol) . At that time, the argon line was replaced with a hydrogen balloon and the reaction was stirred overnight. The mixture was filtered through a bed of Celite ® and the filtrate was concentrated under reduced pressure. The solid residue was triturated with ethyl ether / hexanes to provide the desired product. (1.2 g, 67%). TLC (4% acetone / 96% methylene chloride) R f 0.09; 1 H NMR (DMSO- d 6) δ 2.67-2.83 (m, 4H), 4.83 (sa, 2H), 6.45 (d, J = 8.2Hz , 2H), 6.84 (d, J = 8.2Hz, 2H), 7.20 (d, J = 6Hz, 2H), 8.41 (d, J = 6Hz, 2H).

Intermediate B7

Preparation of 3-fluoro-4- (4-pyridinylsulfanyl) aniline

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Stage one

Preparation of 4 - [(2-fluoro-4-nitrophenyl) sulfanyl] pyridine

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A solution of 4-mercaptopyridine (4.2 g, 35.6 mmol), 3,4-difluoronitrobenzene (5.7 g, 35.7 mmol), and potassium carbonate (12.4 g, 89.7 mmol) in anhydrous DMF (40 ml) it was stirred at 40 ° C and under argon for 3 hours. TLC showed the complete reaction. The mixture was diluted with ethyl acetate (100 ml) and water (100 ml) and the aqueous phase was reextracted with ethyl acetate (2 x 100 ml). The organic phases were washed with a saturated NaCl solution (100 ml) dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by column chromatography with 50% ethyl acetate / 50% hexanes. It provided the desired product in the form of a yellow solid (6.3 g, 71%). TLC (50% EtOAc / 50% hexane). R f 0.53; 1 H NMR (DMSO- d 6 ) δ 7.27 (dd, J = 0.76, 4.2 Hz, 2H), 7.78 (dt, J = 0.76, 7, 2 Hz, 1H), 8.11 - 8.15 (m, 1H), 8.28 - 8.33 (m, 1H), 8.5 (dd, J = 1.4, 4.6 Hz, 2H ), MS [M + H] + = 251.

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Stage 2

Preparation of 3-fluoro-4- (4-pyridinylsulfanyl) aniline

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A suspension of 3-fluoro-4-pyridinylthio) nitrobenzene (6.3 g, 25.2 mmol), iron powder (6.0 g, 107.4 mmol), acetic acid (100 ml), and water (1 ml) were stirred at room temperature overnight. The mixture was diluted with Et2O (100 ml) and water (100 ml). The aqueous phase was adjusted to pH 5 with a 4 N NaOH solution. The combined organic phases were washed with a saturated aqueous NaCl solution (100 ml), dried (MgSO 4) and concentrated under reduced pressure. The residue was purified by column chromatography with 50% ethyl acetate / 50% hexanes. It provided the desired product in the form of a white solid (4.8 g, 86%). TLC (50% EtOAc / 50% hexane). R f 0.28; 1 H NMR (DMSO-d 6) δ 6.04 (sa, 2H), 6.47-6.51 (m, 2H), 6.91 (d, J = 6.1 Hz , 2H), 7.22 (t, J = 8.4 Hz, 1H), 8.30 (d, J = 6.4 Hz, 2H).

Using procedures similar to those described for the preparation of Intermediates B1 - B7, it is also They prepared the following additional compounds:

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TABLE 2 Intermediates B of arylamine

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C. Preparation of the examples of the invention Example 1 Preparation of N - (2-amino-6-methyl-4-piyrimidinyl) - N - [3-fluoro-4- (4-pyridinylsulfanyl) phenyl] amine

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A suspension of 2-amino-4-chloro-6-methylpyrimidine (Intermediate A7, 0.2 g, 1.3 mmol), 3-fluoro-4- (4-pyridinylthio) aniline (Intermediate B7, 0.3 g, 1.3 mmol), and K 2 CO 3 (0.2 g, 1.3 mmol) in o- xylene (1.3 ml) was heated at 100 ° C in a 5 ml reaction vial throughout the night. The reaction mixture was diluted with MeOH and coated on silica and purified by MPLC (Biotage) with 5% -7% MeOH in CH2Cl2. It provided 74 mg of product (18% yield). TLC (6% MeOH / 94% CH 2 Cl 2) R f 0.29; MS ES 328 [M + H] +; 1 H NMR (DMSO- d 6 ) δ 2.12 (s, 3H), 5.92 (s, 1H), 6.38 (sa, 2H), 6.96 (d, J = 5.1 Hz, 2H), 7.39 - 7.52 (m, 2H), 8.26 (d, J = 11.9 Hz, 1H), 8.33 (d, J = 4.8 Hz , 2H), 9.55 (sa, 1H).

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Example 2 Preparation of 6-ethyl- N 4- [3-fluoro-4- (4-pyridinylsulfanyl) phenyl] -2,4-pyrimidinadiamine

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2-Amino-4-chloro-6-ethylpyrimidine (Intermediate A8, 55.1 mg, 0.25 mmol) and Intermediate B7 (39.4 mg, 0.25 mmol) were suspended in aqueous 0.01 M HCl (500 \ mul). The mixture was refluxed for 6 hours. The reaction was cooled to room temperature and the solvent was evaporated in vacuo. The residue was purified by reverse phase chromatography on a YMC Pack-pro C18 column (trade mark) eluting with acetonitrile / H2O (gradient 10:90-90:10). The compound was purified again by TLC preparation eluting with CH2Cl2-MeOH (90:10). Desired compound (2.9 mg, 0.0085 mmol; 34% yield); 1 H NMR (Methanol- d 4) 8.16 (dd, J = 1.7, 4.7, 2H), 8.00-8.04 (m, 1H), 7.37 (m, 2H), 6.93 (dd, J = 1.8, 4.9, 2H), 5.91 (s, 1H), 2.39 (q, J = 7.7, 2H), 1.13 (t, J = 7.5, 3H); ES MS [M + H] + = 342; TLC (CH 2 Cl 2 - MeOH, 90:10); R_ {f} = 0.48.

Examples 3-26

Using the above procedures, the following examples of pyridines were synthesized and summarized in the Table 3.

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Selecting combinations of Intermediates appropriate A1-A21 with Intermediates B1-B17, a diversity of products similarly and are described in the Examples 27-31.

Example 27 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - {4 - [(4-pyridinylmethyl) sulfanyl] phenyl} amine

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Prepared in 34% yield from Intermediate A7 and B8: TLC (7% MeOH in CH 2 Cl 2) R f 0.36; MS (ES) 324 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.09 (s, 3H), 4.12 (s, 2H), 5.87 (s, 1H), 6.33 (sa, 2H ), 7.19 (d, J = 8.5 Hz, 2H), 7.23 (d, J = 5.8 Hz, 2H), 7.64 (d, J = 8.5 Hz, 2H), 8.43 (d, J = 5.3 Hz, 2H), 9.20 (sa, 1H).

Example 28 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - {4 - [(4-pyridinylsulfanyl) methyl] phenylyl} amine

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Prepared in 6% yield from Intermediate A7 and B8: TLC (7% MeOH in CH 2 Cl 2) R f 0.38; MS (ES) 342 [M + H] +; 1 H NMR (DMSO-d 6) 8 2.06 (s, 3H), 4.30 (s, 2H), 5.84 (s, 1H), 6.13 (sa, 2H) , 7.27-7.31 (m, 4H), 7.63 (d, J = 7.9 Hz, 2H), 8.33 (d, J = 6.1Hz, 2H), 8.99 (sa , 1 HOUR).

Example 29 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - {4- [2- (4-pyridinyl) ethyl] phenyl} amine

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Prepared in 30% yield from Intermediate A7 and B6: TLC (8% MeOH in CH 2 Cl 2) R f 0.34; MS (ES) 306 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.06 (s, 3H), 2.83-2.87 (m, 4H), 5.82 (s, 1H), 6.09 (sa, 2H), 7.07 (d, J = 8.5 Hz, 2H), 7.21 (d, J = 5.8 Hz, 2H), 7.54 (d, J = 8.5 Hz , 2H), 8.41 (d, J = 6.2 Hz, 2H), 8.87 (s, 1H).

Example 30 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [4- (4-pyridinylsulfanyl) -3- (trifluoromethyl) phenyl] amine

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Prepared in 1.2% yield from A7 and B9: TLC (7% MeOH in CH 2 Cl 2) R f 0.39; MS (ES) 378 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.03 (s, 3H), 5.94 (s, 1H), 6.33 (sa, 2H), 6.91 (d, J = 6.5 Hz, 2H), 7.64 (d, J = 8.9 Hz, 1H), 8.19 (d, J = 2.2 Hz, 1H), 8.33 (d, J = 5 , 9 Hz, 2H), 8.37 (dd, J = 2.1, 8.6 Hz, 1H), 9.66 (s, 1H).

Example 31 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [4- (5-isoquinolinyloxy) phenyl] amine

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Prepared in 30% yield from A7 and B10: TLC (6% MeOH in CH 2 Cl 2) R f 0.39; MS (ES) 344 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.08 (s, 3H), 5.85 (s, 1H), 6.11 (sa, 2H), 7.02-7.08 (m, 3H), 7.58 (t, J = 8.1 Hz, 1H), 7.74 (d, J = 8.6 Hz, 2H), 7.84 (d, J = 8.2 Hz , 1H), 7.98 (d, J = 5.8 Hz, 1H), 8.54 (d, J = 5.9 Hz, 1H), 9.03 (sa, 1H), 9.35 (sa , 1 HOUR).

Example 32 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [3-fluoro-4- (5-isoquinolinyloxy) phenyl] amine

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A suspension of 2-amino-4-chloro-6-methylpyrimidine (Intermediate A7, 0.14 g, 1.0 mmol), 3-fluoro-4- (5-isoquinolin-oxy) aniline (Intermediate B10, 0.25 g, 1.0 mmol), and HCl (0.1 ml) in H2O (1.0 ml) was heated at 70 ° C in a 5 ml reaction vial overnight. The reaction mixture was diluted with MeOH, treated with saturated NaHCO 3, and coated on silica and purified by MPLC (Biotage) with 5% MeOH in CH 2 Cl 2. It provided 52 mg of product (14% yield). TLC (6% MeOH / 94% CH 2 Cl 2) R f 0.45; MS (ES) 362 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.10 (s, 3H), 5.88 (s, 1H), 6.29 (sa, 2H), 6.93 (d, J = 7.9 Hz, 1H), 7.22 (t, J = 8.9 Hz, 1H), 7.34 (dd, J = 1.7, 8.9 Hz; 1H), 7.55 (t , J = 8 , 1 Hz, 1H), 7.82 (d, J = 8.3 Hz, 1H), 8.08 (d, J = 5.6 Hz, 1H), 8.21 (dd, J = 2.6, 14.2 Hz, 1H), 8.58 (d, J = 5.9 Hz, 1H), 9.30 (sa, 1H), 9.36 (s, 1H).

Using the procedure described above for Example 32, Examples 33-41 were prepared in a manner Similary.

Example 33 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [3,5-dichloro-4- (4-pyridinylsulfanyl) phenyl] amine

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Prepared in 10% yield from A7 and B14: TLC (5% MeOH in CH 2 Cl 2) R f 0.14; MS (ES) 378 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.14 (s, 3H), 5.92 (s, 1H), 6.49 (sa, 2H), 6.93 (d, J = 6.1 Hz, 2H), 8.13 (s, 2H), 8.35 (d, J = 6.2 Hz, 2H), 9.63 (sa, 1H).

Example 34 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [3,5-dichloro-4- (5-isoquinolinyloxy) phenyl] amine

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Prepared in 58% yield from A7 and B12: TLC (70% EtOAc / 30% hexanes) R f 0.18; MS (ES) 412 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.13 (s, 3H), 5.89 (s, 1H), 6.38 (sa, 2H), 6.73 (d, J = 7.6 Hz, 1H), 7.52 (t, J = 7.9 Hz, 1H), 7.82 (d, J = 8.2 Hz, 1H), 8.05 (s, 2H), 8.16 (d, J = 5.8 Hz, 1H), 8.61 (d, J = 5.9 Hz, 1H), 9.36 (s, 1H), 9.42 (s, 1H).

Example 35 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [6- (4-pyridinylsulfani) -3-pyridinyl] amine

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Prepared in 16% yield from A7 and B13: TLC (6% MeOH in CH 2 Cl 2) R f 0.16; MS (ES) 311 [M + H] +; 1 H NMR (DMSO-d 6) δ 2.11 (s, 3H), 5.91 (s, 1H), 6.34 (sa, 2H), 7.15 (d, J = 6.2 Hz, 2H), 7.48 (d, J = 8.8 Hz, 1H), 8.30 (dd, J = 2.5, 8.4 Hz, 1H), 8.38 (d , J = 6.1 Hz, 2H), 9.00 (d, J = 2.4 Hz, 1H), 9.45 (sa, 1H).

Example 36 Preparation of N - (2-amino-6-phenyl-4-pyrimidinyl) - N - [4- (4-piidinylsulfanyl) phenyl] amine

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Prepared in 65% yield from A17: TLC (4% MeOH in CH 2 Cl 2) R f 0.22; MS (ES) 372 [M + H] +; 1 H NMR (DMSO-d 6) δ 6.46 (sa, 2H), 6.55 (s, 1H), 6.96 (d, J = 4.8 Hz, 2H), 7.46-7.49 (m, 5H), 7.91-8.00 (m, 4H), 8.32 (d, J = 4.9 Hz, 2H), 9.57 (sa, 1H) .

Example 37 Preparation of N - [2-amino-6- (3-pyridinyl) -4-pyrimidinyl] - N - [3-fluoro-4- (4-pyridinylsulfanyl) phenyl] amine

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Prepared in 45% yield. TLC (4% MeOH in CH 2 Cl 2) R f 0.27; MS (ES) 391 [M + H] +; 1 H NMR (DMSO-d 6) δ 6.58 (s, 1H), 6.70 (sa, 2H), 6.99 (d, J = 6.4 Hz, 2H), 7.46-7.57 (m, 3H), 8.23-8.36 (m, 4H), 8.65 (d, J = 4.4 Hz, 1H), 9.09 (s, 1H) , 9.86 (sa, 1 H).

Example 38 Preparation of N - [2-amino-6- (4-pyridinyl) -4-pyridinyl] - N - [3-fluoro-4- (4-pyridinylsulfanyl) phenyl] amine

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Prepared in 22% yield from A2 and B7: TLC (6% MeOH in CH 2 Cl 2) R f 0.32; MS (ES) 391 [M + H] +; 1 H NMR (DMSO-d 6) δ 6.63 (s, 1H), 6.74 (sa, 2H), 6.99 (d, J = 5.8 Hz, 2H), 7.46-7.58 (m, 2H), 7.85 (d, J = 5.8 Hz, 2H), 8.31-8.36 (m, 3H), 6.70 (d, J = 4.3 Hz, 2H), 9.92 (sa, 1H).

Example 39 Preparation of N - [2-amino-6- (4-pyridinyl) 4-pyrimidinyl] - N - [3,5-dichloro-4- (4-pyridinylsulfanyl) phenyl] amine

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Prepared in 0.4% yield from A2 and B14: TLC (4% MeOH in CH 2 Cl 2) R f 0.15; 1 H NMR (DMSO-d 6) δ? 6.60 (s, 1H), 6.82 (sa, 2H), 6.95 (d, J = 5.9 Hz, 2H) , 7.85 (d, J = 5.9 Hz, 2H), 8.18 (s, 2H), 8.36 (d, J = 3.8 Hz, 2H), 8.71 (d, J = 4.7 Hz, 2H), 9.99 (sa, 1H).

Example 40 Preparation of N- [2-amino-6- (3-pyridinyl) -4-pyrimidinyl] - N - [3-fluoro-4- (5-isoquinolinyloxy) phenyl] amine

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Prepared in 54% yield from A18 and B11: TLC (5% MeOH in CH 2 Cl 2) R f 0.34; MS (ES) 425 [M + H] +; 1 H NMR (DMSO-d 6) δ 6.53 (s, 1H), 6.59 (sa, 2H), 6.96 (d, J = 7.4 Hz, 1H), 7.26 (t, J = 9.6 Hz, 1H), 7.38-7.59 (m, 3H), 7.83 (d, J = 8.2 Hz, 1H), 8.09 (d , J = 5.7 Hz, 1H), 8.23-8.31 (m, 2H), 8.58-8.65 (m, 2H), 9.09 (sa, 1H), 9.37 ( sa, 1H), 9.61 (sa, 1H).

Example 41 Preparation of N - [2-amino-6- (2-pyridinyl) 4-pyrimidinyl] - N - [3-fluoro-4- (4-pyridinylsulfaninl) phenyl] amine

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Prepared in 60% yield from A19 and B7: TLC (50% EtOAc / 50% hexanes) R f 0.14; MS (ES) 391 [M + H] +; 1 H NMR (DMSO-d 6) δ 6.63 (sa, 2H), 6.98 (d, J = 6.6 Hz, 2H), 7.17 (s, 1H), 7.45 - 7.57 (m, 3H), 7.93 (dt, J = 1.9.7.7 Hz, 1H), 8.23 - 8.37 (m, 4H), 8.65 - 8.67 (m, 1 H), 9.90 (sa, 1 H).

Example 42 Preparation of N - (2-amino-6-ethyl-4-pyrimidinyl) - N - [4- (4-pyridinylsulfanyl) phenyl] amine

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A mixture of Intermediate B5 (50.6 mg, 0.250 mmol) and 2-amino-4- ethyl-6-chloropyrimidine (Intermediate A8, 39.4 mg, 0.25 mmol) in aqueous 0.01 M HCl (500 ml ) was refluxed for 6 hours. The reaction was cooled to room temperature and the solvent was evaporated in vacuo. The residue was purified by reverse phase chromatography on a YMC Pack-pro ® column eluting with acetonitrile / H2O (gradient 10:90-90:10) to give the desired compound (13.0 mg, 0.040 mmol; 16% yield); mp = 181 ° C - 186 ° C; ES MS [M + H] + = 324; TLC (CH 2 Cl 2 -MeOH, 95: 5); R f = 0.41.

Example 43 Preparation of N - (2-amino-6-chloro-4-pyrimidinyl) - N - [3-fluoro-4- (4-pyridinylsulfanyl) phenyl] amine

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2-Amino-4.6 dichloropyrimidine (A21, 12 mmol) and 3-fluoro-4- (4-pyridinylthio) -aniline (B7, 12 mmol) were suspended in water (150 ml) and treated with 10 drops of concentrated hydrochloric acid. The mixture was stirred at 100 ° C. All night long. The clear solution was neutralized. then with ammonium hydroxide. The yellow precipitated product filtered, washed with water, and purified by chromatography on column with 1% MeOH - 3% in CH2Cl2 to give the desired product in the form of a white solid (1.98 g, 47%).

Example 44 Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [2,5-difluoro-4- (4-pyridinylsulfanyl) phenyl] amine

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The compound was prepared using a similar procedure used for the preparation of Example 1 (described above) from A7 and B15: HPLC / MS: [M + H] + 346.1 m / z . Retention time (HPLC / MS) = 1.39 min.

Example 45 Preparation of N - (2-amino-6-chloro-4-pyrimidinyl) - N - (3,5-dichloro-4- (4-pyridinylsulfanyl) phenyl] amine

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The compound was prepared using a similar procedure used for the preparation of Example 43 (described above) from A21 and B14: HPLC / MS: [M + H] + 399.0 m / z . Retention time (HPLC / MS) = 3.02 min.

Example 46 Preparation of N - (2-amino-6-chloro-4-pyrimidinyl) - N - [3-fluoro-4- (5-isoquinolinyloxy) phenyl] amine

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The compound was prepared using a similar procedure used for the preparation of Example 43 (described above) from A21 and B11: HPLC / MS: [M + H] + 382.1 m / z . Retention time (HPLC / MS) = 2.91 min.

Examples 47-49

Using procedures similar to the examples above and using the appropriate Intermediates A and B, were prepared The following examples similarly:

Example 47

Preparation of N - (2-amino-6-methyl-4-pyrimidinyl) - N - [1- (4-pyridinyl) -1 H -indole-5-yl] amine

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Prepared by means of the reaction of Intermediate A7 and B1.

Example 48

Preparation of N - (2-anilino-6-methyl-4-pyrimidinyl) - N - [1- (4-pyridinyl) -1H-indol-5-yl] amine

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Prepared by means of the reaction of Intermediate A16 and B1.

Example 49

Preparation of N - (2-anilino-6-methyl-4-pyrimidinyl) - N [4-fluoro-3- (4-pyridinylsulfanyl) phenyl] amine

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Prepared by means of the reaction of Intermediate A16 and B7.

Rho kinase biochemical assay

Activity criteria of ROCK-1: 0 no effect (<40% inhibition), 1 effect (> 40% inhibition). Trial tests for the ROCK-1 phosphorylation inhibition of MBP (Basic Myelin Protein). The reaction (100 µl of final volume) is carried out in 96-well plates of Polypropylene in 50 mM HEPES buffer pH 7.5 containing MgCl 2 mM and DTT 1 mM. For each well gstROCK1 (0.25 \ mug of gstROCK1 of BAYER DRT) is combined with MBP (1 mug) in reaction buffer (combined volume 70 µl). Inhibitors are added (5 µl of 20 x conc. in 40% DMSO) to each well to provide a dose response range at 8 points from 1.0 µM to 5 nM. The reaction is started by adding 25 µL of ATP (4x = 12 µM) in reaction buffer containing 0.8 µCi of 33 P gamma-ATP (4x) to provide a concentration 3 µM cold and 0.2 µCi hot ATP finish. The plates are incubated for 1 hour at room temperature stopping the reaction by adding 7 µl of 1 N HCl. MBP Radiolabelled was transferred to P30 filter units (EG&G Wallac), washed in 1% phosphoric acid followed by brief washings in water. Then the filter units were dried and incorporation of 33 P was detected by scintillation counting liquid. The background incorporation of 33 P is determined by the autophosphorylation of ROCK1 without MBP. The data is expressed as percentage inhibition:% inhibition = 1 - ((cpm with inhibitor - of background) / (cpm without inhibitor - background)) * 100.

Claims (5)

1. A compound of the formula

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84 85 86 87 88 89 90 91 92 93 94 95 96 2. Use of a compound as defined in the claim 1, for the manufacture of a drug useful for the treatment of an indication mediated by Rho kinase. 3. Use according to claim 2 for the manufacture of a drug for humans. 4. Use of a compound as defined in the claim 1, for the manufacture of a drug useful for the treatment of hypertension, atherosclerosis, restenosis, cerebral ischemia, cerebral vasospasm, degeneration Neural, spinal cord injury, breast cancer, colon, prostate, ovaries, brain or lung, thrombotic disorders, the asthma, glaucoma, osteoporosis or erectile dysfunction. 5. Use according to claim 4 for the manufacture of a drug for humans.